Department of Molecular Systems Biology, UFZ, Helmholtz-Centre for Environmental Research, Permoserstraße 15, 04318 Leipzig, Germany.
Department of Medicine, University of Leipzig, Liebigstraße 27b, 04103 Leipzig, Germany.
Molecules. 2020 Feb 11;25(4):775. doi: 10.3390/molecules25040775.
Adipokines and apolipoproteins are key regulators and potential biomarkers in obesity and associated diseases and their quantitative assessment is crucial for functional analyses to understand disease mechanisms. Compared to routinely used ELISAs, multiple reaction monitoring (MRM)-based mass spectrometry allows multiplexing and detection of proteins for which antibodies are not available. Thus, we established an MRM method to quantify 9 adipokines and 10 apolipoproteins in human serum. We optimized sample preparation by depleting the two most abundant serum proteins for improved detectability of low abundant proteins. Intra-day and inter-day imprecision were below 16.5%, demonstrating a high accuracy. In 50 serum samples from participants with either normal weight or obesity, we quantified 8 adipokines and 10 apolipoproteins. Significantly different abundances were observed for five adipokines (adipsin, adiponectin, chemerin, leptin, vaspin) and four apolipoproteins (apo-B100/-C2/-C4/-D) between the body mass index (BMI) groups. Additionally, we applied our MRM assay to serum samples from normal weight children and human adipocyte cell culture supernatants to proof the feasibility for large cohort studies and distinct biological matrices. In summary, this multiplexed assay facilitated the investigation of relationships between adipokines or apolipoproteins and phenotypes or clinical parameters in large cohorts, which may contribute to disease prediction approaches in the future.
脂联素和载脂蛋白是肥胖症及相关疾病的关键调节因子和潜在生物标志物，其定量评估对于功能分析、了解疾病机制至关重要。与常规使用的 ELISA 相比，基于多重反应监测 (MRM) 的质谱法可以对没有抗体的蛋白质进行多重检测和检测。因此，我们建立了一种 MRM 方法，用于定量检测人血清中的 9 种脂联素和 10 种载脂蛋白。我们通过耗尽两种最丰富的血清蛋白来优化样品制备，以提高低丰度蛋白的检测能力。日内和日间精密度均低于 16.5%，表明准确性高。在 50 名体重正常或肥胖的参与者的血清样本中，我们定量检测了 8 种脂联素和 10 种载脂蛋白。在 BMI 组之间，五种脂联素（adipsin、adiponectin、chemerin、leptin、vaspin）和四种载脂蛋白（apo-B100/-C2/-C4/-D）的丰度存在显著差异。此外，我们还将我们的 MRM 测定法应用于体重正常儿童的血清样本和人脂肪细胞培养上清液，以证明其在大型队列研究和不同生物基质中的可行性。总之，这种多重检测方法促进了在大型队列中研究脂联素或载脂蛋白与表型或临床参数之间的关系，这可能有助于未来的疾病预测方法。
